Plant Foods Hum Nutr (2007) 62:139144

DOI 10.1007/s11130-007-0054-8

ORIGINAL PAPER

Chemical Composition of Green Tea (Camellia sinensis)

Infusions Commercialized in Portugal
Mrcia Reto & Maria Eduardo Figueira &
Helder Mota Filipe & Cristina M. M. Almeida

Published online: 27 September 2007

# Springer Science + Business Media, LLC 2007

Abstract To evaluate the potential benefits and risks associated with tea consumption it is important to identify the
constituents of this beverage. Levels of some minerals,
caffeine and catechins in green tea samples commercialized
in Portugal were evaluated. Potassium is the metal present in
larger amount (92151 mg/l). The content of sodium, calcium,
fluoride, aluminium, manganese and iron were 3569, 1.9
3.5, 0.802.0, 1.02.2, 0.521.9, 0.0200.128 mg/l, respectively. Chromium and selenium were not detected. The
resulting data showed considerable variability in catechins
content. The levels of epigallocatechin gallate (EGCG) ranged
from 117 to 442 mg/l, epicatechin 3-gallate (EGC) from 203
to 471 mg/l, epigallocatechin (ECG) from 16.9 to 150 mg/l,
epicatechin (EC) from 25 to 81 mg/l and catechin (C) from
9.03 to 115 mg/l. Caffeine contents in the green tea infusions
studied were between 141338 mg/l. Green tea infusions
provide significant amounts of catechins and could be an
important source of some minerals.
Keywords Caffeine . Catechins . Fluoride . Green tea .
Minerals
Abbreviations
C
(+) catechin
EC
() epicatechin

M. Reto
Faculdade de Farmcia da Universidade de Lisboa,
Lisbon, Portugal
M. E. Figueira : H. M. Filipe : C. M. M. Almeida (*)
Unidade Farmacologia e Farmacotoxicologia,
Faculdade de Farmcia da Universidade de Lisboa,
Av. das Foras Armadas,
1600-083 Lisbon, Portugal
e-mail: calmeida@ff.ul.pt

ECG
EGC
EGCG
HPLC
M4
M6

()epicatechin gallate
()epigallocatechin
()epigallocatechin gallate
High-performance liquid chromatography
()-5-(3,4, 5-trihydroxyphenyl)-gammavalerolactone
()-5-(3, 4-dihydroxyphenyl)-gammavalerolactone

Introduction
Tea is an infusion of Camellia sinensis leaves. Native from
China it is today the most widely consumed beverage in the
world, aside water.
During the last decades, the beneficial health effects of
green tea have been demonstrated in a number of studies.
Tea drinking is associated with reduction of serum
cholesterol, prevention of low density lipoprotein (LDL)
oxidation and a decreased risk of cardiovascular disease
and cancer [1, 2]. Due to these facts the popularity of this
beverage has grown all over the world.
The chemistry of tea is complex: polyphenols, alkaloids,
amino acids, glucides, proteins, volatile compounds, minerals and trace elements [3]. Polyphenols are the most
biologically active group of tea components which have
antioxidative, antimutagenic and anticarcinogenic effects
[4, 5]. But there are other compounds in green tea with
interest for human health like fluoride, caffeine, minerals
and trace elements like, chromium and manganese [68].
The regular consumption of tea can contribute to the daily
dietary requirements of some of these elements [9]. Due to
the importance of minerals in tea several studies have been
carried out in order to determine their levels in tea leaves
and infusions [1014].

140

To evaluate the nutritional contribution and the intoxication risk related with green tea consumption it is
important to quantify micronutrients and toxic elements
levels in this beverage. Therefore aluminium, chromium,
selenium, manganese, iron, sodium, potassium, calcium,
fluoride, caffeine and catechins were determined in green
tea infusions available in Portugal.

Materials and Methods

Tea Samples
In Portugal the consumption of green tea is new. The
market is growing and there are only a few brands of green
tea. For this study were selected the most representative
green tea brands accordingly to the information of the
specialty shops. These tea samples were purchased from
several markets and herbalists in Lisbon region. Six of them
were sold in tea bags and the other three as loose-leaf tea.
The samples were identified by the acronyms G, S, T, TN,
TV, F, B, M and T. Each tea was analysed in triplicate using
separate infusions.
The infusion time and ratio sample/water volume were
selected based on the label instructions of the analyzed
samples (infusion of a tea bag in a cup of boiling water
(250 ml) and a brewing time from 510 min) in order to
simulate the conditions used by the consumers at home.
A mass of 1.5 g of tea leaf (or tea leaf from a tea bag)
was weighed in a beaker and 250 ml of boiling ultra-pure
water were added. After 10 min, tea infusions were filtered
into a standard flask of 250 ml and diluted to the mark with
ultra-pure water. All the infusions were analyzed in the day
of the preparation.
Methods
The fluoride concentration in green tea infusion was
determined using a digital pH meter, with a fluoride
selective electrode (Mettler Toledo, Switzerland) and a
Ag/AgCl as reference electrode (Mettler Toledo, Switzerland) [15]. For calcium determination the EDTA titrimetric
method was performed accordingly with the described by
Clesceri et al. [15].
Potassium and sodium were analyzed by flame photometric method using a flame photometer 400 flame
photometer Corning 450 (Corning Limited, Essex, England) [15].
Aluminium, chromium, selenium, manganese and iron
were determined by electrothermal atomic absorption
spectrometry (EAAS) with Perkin Elmer Zeeman Atomic
Spectrometer 4110ZL and Autosampler AS-72 from Perkin
Elmer (Norwalk, Connecticut). The appropriate conditions

Plant Foods Hum Nutr (2007) 62:139144

for each metal were used [15]. The material used to prepare
the infusions was washed with nitric acid and rinsed with
ultra-pure water. The samples were analyzed after a pretreatment with concentrate HNO3. 250 ml of green tea
infusion were transferred to a beaker with boiling chips. In
a hood, 5 ml of concentrate HNO3 were added. The beaker
was covered with a ribbed watch glass to minimise
contamination. After a slow boil, the solution was evaporated, on a hot plate (DataPlateDigital, Biomolecular Inc.
from Cole-Parmer International, Vernon Hills, U.S.A.), to
about 150 ml. Another 5 ml of concentrate HNO3 were
added until digestion was completed, as showed by a light
yellow clear solution. The solution was brought to the
lowest volume possible before precipitation (about 70
90 ml). The digested solution was transferred to a 100 ml
volumetric flask and diluted to the mark with ultra-pure
water. A reagent blank was prepared with the same acid and
subjected to the same digestion procedure as the sample to
correct impurities present in acid and reagent water.
The method proposed by Fernandez et al. [16] was used
for the simultaneous determination of catechins and
caffeine in the tea infusions. A high-performance liquid
chromatograph system equipped with a binary LC-6A
pump controlled by a SCL-6B controller, a ultra violet
detector model SPD-M10A DAD and a LC workstation
(class LC-10 verso1) integrator all from Shimadzu (Kyoto,
Japan) and a Lichrosper 100 RP-18 (5 m), 250 mm
4 mm column (Ecopack, Merck) were used. The tea
infusions were filtered through a 0.45 m filter and then
20 L were injected for HPLC analysis.
Methods Validation
Accuracy was validated by recovery assays. Recoveries
were calculated as the difference between the amount of
analyte present in spiked and non-spiked samples divided
by the amount of added analyte [17].
The linear range of each method was assessed by
analyzing several solutions containing standards in different
concentration levels.
Ten solutions containing the target compounds at the
lowest concentration of linearity range were analyzed and
the standard deviations (SD) were determined. The values
of limit of detection (LOD) and limit of quantification
(LOQ) were calculated using the formula (3SD) and (10
SD) respectively [18].
The precision of each method was evaluated by carrying
out ten replicate analyses of a standard solution, with a
concentration equivalent to the lower limit of the linearity
range. The relative standard deviation (RSD) should be
lower than 10% accordingly with the requirements of
Portuguese law for water for human consumption (DL
243/01, 5th September).

Plant Foods Hum Nutr (2007) 62:139144

141

Results and Discussion

The linearity range, precision, recovery and limits of
detection (LOD) and quantification (LOQ) obtained are
shown in Table 1.
The results showed, for the majority of methods, a good
precision under the analytical conditions used since the
relative standard deviations are lower than 10%. Good
recoveries between 80115% were obtained.
The contents of the analyzed elements were determined
in the tea samples. The results are summarized in Table 2.
Potassium was the element with higher content in all
samples analyzed ranging from 92151 mg/l. These concentrations were lower than those reported by Powell et al. [7]
(221 mg/l), Matsuura et al. [11] (246 mg/l) and Fernndez et

al. [14] (94259 mg/l). The levels of sodium in the nine

infusions were much higher (3669 mg/l) than those
described in the literature [7, 11, 14]. Different concentrations of this element should be expected in the infusions
since sodium is a highly extractable element and its content
in green tea leaves can range from 33.9 mg/kg to 1760 mg/kg.
[12]. The conditions under which the tea infusions were
prepared are other possible explanation for the observed
differences. Recent research suggests that the Na/K ratio is
important to control blood pressure. A ratio of less than one
is recommended. Since the tea samples analyzed have at least
two or three times as much potassium as sodium, their
consumption may be useful for lowering blood pressure.
Calcium is the most common mineral in the body. The
obtained calcium levels in tea leaves ranged from 1.9

Table 1 Linearity, precision, recoveries and detection and quantification limits of the different methods
Compound

Method

Calcium

EDTA Titrimetric

Sodium

Flame emission
photometric
method

Fluoride

Selective
electrode

R2

Not applicable
5

Potassium

Linearity range
(mg/L)

1.0-10

LOD
(mg/L)

LOQ
(mg/L)

RSD
%
(N=10)

Recovery
(%)
(N=3)

0.5

1.7

1.8

80-110

0.30

1.0

10

0.995

96-107

10-70

0.995

13

0.50-5.0

0.995

0.04

0.14

3.0

96-100

15

0.04-10

0.997

0.07

0.25

13

103-110

0.994

1.2 x10-3

4.0 x10-3

3.0

100-115

Aluminium

Chromium

0.6 x10-3 - 40 x10-3

0.997

0.6 x10-3

2.0 x10-3

4.0

100-115

1.0 x10-3 - 70 x10-3

0.999

1.2 x10-3

4.0 x10-3

4.0

100-115

Manganese

1.0 x10-3 - 40 x10-3

0.999

1.2 x10-3

4.0 x10-3

3.0

100-115

Selenium

0.4 x10-3 - 10 x10-3

0.991

0.36 x10-3

1.2 x10-3

2.0

90-100

Caffeine

75-500

0.999

6.2

20

1.8

106-114

EGC

10

10-500

0.999

1.9

6.4

1.8

93-96

13

2.5-500

0.999

0.68

2.2

1.8

86-118

12

2.5-500

0.999

0.69

2.3

1.5

90-117

12

5-500

0.999

0.45

1.5

2.9

92-108

10-150

0.999
2.2

7.4

2.5

90-118

75-300

0.999

Iron

EC
EGCG

Electrothermal
atomic absorption
spectrometry

HPLC-UV

1.0x10-3 - 40 x10-3

ECG

N Number of experimental points

142

Plant Foods Hum Nutr (2007) 62:139144

Table 2 Content of the different elements in the green tea infusions (N=3)
Green tea

Ca

Na

Al

Mn

Cr

Fe

Se

TV
TN
T
D
F
B
M
S
G

2.90.2
1.90.2
2.60.2
3.50.2
3.40.2
3.00.2
2.20.2
2.10.2
1.90.2

1131
10811
1202
1148
1113
12118
921
1513
1031

550
512
471
455
521
433
351
521
691

1.40.1
1.10.1
1.30.1
1.20.1
1.00.1
1.10.1
0.900.1
0.800.1
2.00.2

1.60.1
1.900.1
1.70.3
1.80.3
1.90.0
2.20.3
2.10.1
1.00.1
1.40.4

1.90.2
1.00.2
1.80.3
1.50.1
1.40.2
1.30.1
1.40.2
1.50.1
0.520.1

n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.

0.0300.003
0.0240.001
0.0450.002
0.0420.003
0.0400.001
0.0200.001
0.1280.007
0.0300.001
0.0220.003

n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.

Units Standard deviation (mg/lSD)

n.d. not detected

3.5 mg/l. These values are consistent with data reported

previously [11].
The anticariogenic effect of fluoride is well known. The
fluoride content of most foods is low (less than 0.05 mg/100 g).
Camellia sinensis plant takes fluoride from soil and accumulates it in the leaves [10]. The fluoride levels of the analyzed
samples were between 0.80 and 2.0 mg/l, in agreement with
previous findings [10, 13]. Simpson et al. [19] showed that
34% of tea fluoride was retained in the oral cavity, suggesting
that this drink is an effective vehicle for delivering fluoride in
the oral cavity helping to prevent dental decay. Some concern
has been raised over excessive intakes of fluoride causing
fluorosis (mottling and decolouration of the teeth). In areas
with fluorinated water supply the consumption of several cups
of green tea per day could lead to an excessive ingestion of
fluoride.
Analysis of variance of the results of Na, K, Ca and F
showed statistically significant differences in content of
these elements among the different teas (p<0.05).
The analysis of Al, Mn, Cr, Fe by EAAS showed that
these trace elements are present in green tea infusions over
a wide range of concentrations (Table 2). Among them,
selenium is the element present with a lower concentration
(<1.2 g/l). In most tea leaves the level of this element is

0.1 g/g but Xie et al. [9] detected higher concentrations

(7.5 g/g) in some tea samples produced in a region with
selenium rich soils.
Tea contains only trace amounts of iron (0.043 and 0.03
0.17 mg/l) [11, 14]. In the present study iron levels were
between 0.0200.128 mg/l. The bioavailability of this
element is influenced by polyphenols found in tea that
can markedly inhibit the absorption of nonheme iron [20].
Tea is an important source of manganese and chromium
which are essential nutrients required for normal metabolism [69]. Mean chromium intakes from food and water
range from 52 to 943 g/day. Chromium was not detected
in the green tea samples analyzed and for this reason the
contribution of these infusions for the daily intake of this
element is insignificant.
Manganese is present in small amounts in foods [9]. The
levels of manganese detected (0.521.9 mg/l) were lower
than those described in other studies [11, 14]. According to
Powell et al. [7] only 40% of the manganese is in a
potentially bioavailable form, so a cup of green tea (225 ml)
may contribute with 10% of the average daily dietary intake
of this metal.
Tea plants take up a substantial amount of aluminium
however this beverage is not among the dietary sources of

Table 3 Concentrations of caffeine and catechins in green tea infusions by HPLC-UV (the compounds were ordinate by their retention time)
Green tea

EGC

Caffeine

EC

EGCG

ECG

Total catechins

TV
TN
T
D
F
B
M
S
G

25919
35814
35010
30113
4710
4511
20519
45112
20311

93.20.1
1152
31.44
31.21
40.52
38.66
9.491
22.62
9.031

2361
3298
2695
25117
3387
3126
2071
2495
1412

813
713
445
2514
583
546
322
564
341

2336
33015
3018
32117
44210
38710
2184
3051
1178

14517
1506
77.88
72.82
1155
76.514
45.414
44.611
16.91

8128
10238
8617
75411
11275
10067
5107
8794
3984

Units Standard deviation (mg/lSD); N=3

Plant Foods Hum Nutr (2007) 62:139144

aluminium associated with an increased risk of Alzheimers

disease [1]. The content of aluminium detected (1.0
2.1 mg/l) was lower than that obtained by Fernndez et
al. [14]. This element, like iron, has poor biovailability due
to the actions of polyphenols, which avidly bind to trivalent
metal and prevent their intestinal absorption.
A considerable variability in metal and trace elements
content was observed in the green tea samples. Although
present in the infusions not all minerals are available in our
organisms after the consumption of a cup of tea since
absorption of most minerals is very variable and incomplete
[21].
The main classes of polyphenols present in green tea are
flavanols and flavonols. These compounds constitute 16
30% of the dry weight of fresh leaf [22]. Catechins (flavan3-ols) are the predominant form found. Fresh green tea
leaves contain five major catechins: catechin (C), ()epicatechin (EC), ()-epicatechin 3-gallate (ECG), ()epigallocatechin (EGC) and ()-epigallocatechin gallate
(EGCG). These compounds are water-soluble and contribute to bitterness and astringency of green tea. Flavonols
such as quercetin, kaempferol, myricitin and their glycosides can also be found in green tea [23, 24].
The concentrations of catechins and caffeine are presented in Table 3.
The obtained results indicated that in the majority of the
samples the EGCG was the catechin present at the highest
concentration, while catechin showed the lowest level. The
detected values of catechins in the samples analyzed are
similar to those described by Wang et al. [25], except for
catechin (C) and ECG but different from those reported by
Del Rio et al. [24] and Lin et al. [26]. The differences
observed can be related with sample preparation methodologies since different brewing times and ratio leaves/water
were used. The comparison of the results is sometimes
difficult due to the lack of uniformity in the conditions used
to prepare the infusions. The amount of leaf to be used per
amount of water is of critical importance. However there
are other factors to consider like the temperature and the
time of infusion. An enhancement of caffeine and catechins
extraction is observed as the temperature increases. The
catechin content of the tea infusion increases 3040% when
extracted for a period of 10 min. The leaf age is also a very
important factor because old leaves contain less caffeine but
more EGCG and total catechins than young ones [27].
Because individual catechins have different antioxidative and health properties is important to know the
concentration of each one of them in the infusions. Toshi
et al. [28] had observed that the antioxidant activity of the
green tea is higher in the teas that contain higher levels of
EGCG and EGC.
The consumption of a cup of tea may contribute to the
intake of 90230 mg of catechins, so a moderate consump-

143

tion may provide significant amounts of these potential

beneficial compounds. The data available until now do not
allow us to predict the amount of polyphenols that have
physiological activity in humans. In the last years several
works had been conducted in order to study the biological
effects, bioavailability and metabolism of tea polyphenols
in humans. These studies are sometimes discordant and
variability among individuals is also observed. Most of the
ingested catechins are rapidly and extensively metabolized
[()-5-(3,4, 5-trihydroxyphenyl)-gamma-valerolactone
(M4) and ()-5-(3, 4-dihydroxyphenyl)-gamma-valerolactone (M6)]. Substantial amounts of these metabolites are
present in plasma and urine after tea consumption so, these
compounds had been used as biomarkers. However the
pharmokinetics of M4 and M6 requires further clarification
in order to define appropriate dose levels [1].
The levels of caffeine in infusions were between 141
338 mg/l. The detected values are in agreement with
previous findings [25, 26, 27]. For the healthy adult
population moderate daily caffeine intake at a dose level
up to 400 mg/kg (6 mg/kg of body weight) is not associated
with adverse effects [29].
Tea has been studied extensively for its biological action
but there are few data on the catechin content of green tea
infusions. Most of the studies have determined the catechin
contents of fresh leaf after extraction with organic solvents.
This work is a contribution for the chemical characterization of green tea infusions. The obtained results
suggested that there are significative differences in the
chemical composition of green tea brands commercialized
in Portugal. However, further studies are needed in order to
obtain a more representative data.
In conclusion, green tea consumption may contribute to
daily intake of some minerals, such as manganese.
Acknowledgment The authors thank all collaborators of the
following laboratories: Laboratrio de Hidrologia e Anlises Hidrolgicas of FFUL (Faculdade de Farmcia da Universidade de Lisboa)
and LALA (Laboratrio de guas do Litoral Alentejano).

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